The purpose of this study was to investigate whether a six-week, twice weekly resistance training (4 sets at 30% 1-RM until failure) with practical blood flow restriction (BFR) using 7cm wide cuffs with a twist lock placed below the patella is superior to training without BFR (NoBFR) concerning muscle mass and strength gains in calf muscles.A two-group  years, training experience 7.3 (7.0) years; NoBFR n = 9, mean age 28.9 (7.4) years, training experience 7.1 (6.6) years) randomized matched pair design based on initial 1-RM was used to assess the effects on structural and functional adaptations in healthy males .The mean number of repetitions completed per training session across the intervention period was higher in the NoBFR group compared to the BFR group (70 (16) vs. 52 (9), p = 0.002). VAS measured during the first session increased similarly in both groups from first to fourth set (p<0.001). No group effects or time×group interactions were found for CV, MT, LS, and 1-RM. However, there were significant time effects for MT , and 1-RM .LS and CV remained unchanged through training. VAS in both groups were similar, and BFR and NoBFR were equally effective for increasing 1-RM and MT in trained males. However, BFR was more time efficient, due to lesser repetition per training session. In addition to the gluteal and upper leg muscles, the calf muscles have a crucial impact on the sprint and jumping performance, as they develop most of the final push, that accelerates an athlete upwards and forward. In a recently published study, Möck et al. presented medium to strong correlations between the one-repetition maximum 1-RM) standing calf raises and sprint performance 30m) of sport students -RM stand. Moreove0m of spoIn order to achieve muscular adaptations in the sense of muscle hypertrophy and strength gains, it was long assumed that moderate to high resistances had to be overcome. However, recent research indicates that mechanical tension through loading is not the only way to achieve these adaptations . An alteIt can be assumed that, based on previous findings, that the applied training stimulus in the form of classical RT was not optimal for achieving muscular hypertrophy of the calf musculature. Metabolic stress may be more suitable. Research also demonstrates that RT of the calf muscles with BFR is able to increase microvascular filtration capacity . In combTherefore, the aim of the present study was to investigate whether a six-week low-intensity RT with BFR is superior to RT without BFR for inducing muscle mass and strength gains, when performed to volitional muscle failure.A two-group randomized matched-pair design, based on initial 1-RM, was used to assess the effects of a six-week low-intensity BFR or low-intensity conventional calf muscle RT (NoBFR) on structural and functional adaptations. The independent variables in this study consisted of two different training modalities (BFR and NoBFR) and six dependent variables: calf volume, gastrocnemius muscle thickness, leg stiffness, and 1-RM calf raise, as well as a pain assessment during the first training session.Prior to the investigation, ethical clearance was obtained from the IST University of Applied Sciences ethics committee according to the Declaration of Helsinki.The G*Power software  was used in advance to determine necessary sample size via power analysis using a medium effect size  . To accoFor each subject pre- and post-testing was performed at the same time of day. During pre-testing, subjects’ training history was assessed via questionnaire. Subsequently, body mass was measured using electronic weighing scale  and whole calf volume (CV), as an indirect measure for hypertrophy, was calculated using PeroPlus  from a series of vertical and horizontal diameter measurements taken by an optoelectronic device  . Same daMuscle thickness (MT) of the medial gastrocnemius of the right lower leg as a direct measurement for skeletal muscle hypertrophy was measured using B-mode ultrasound  with 8.5-MHz linear probe . Ultrasound is considered a valid measure for muscle thickness changes compared to other techniques such as magnetic resonance imaging or computed tomography ,13. For Subsequently, to warm up, participants completed five minutes low-intensity treadmill running. Leg stiffness (LS) was then assessed using the Optojump photocell system  . For theFollowing five minutes of rest, 1-RM calf raise was conducted using a Smith machine . The participants were positioned with their forefoot on a step and the barbell bar resting on their neck. Maximum ankle plantar flexion with an unloaded bar was measured using a goniometer. Warm-up sets of 10, 5, 3 and 1 repetitions were done using 50, 65, 80 and 90% of the participants estimated 1-RM, interspersed with 1, 2, 3 and 4 minutes of rest respectively . AfterwaAfter pre-testing, all subjects completed 2 sets of 10 repetitions, one with and one without BFR, using 30% of their 1-RM to get familiar with both training modalities. For BFR, a practical approach was utilized , using 7The experiment was conducted from October 2018 to December 2018. The intervention lasted six weeks and consisted of one calf exercise on two non-consecutive days of the week (minimum 48 and maximum 96 hours apart) in addition to the subjects’ usual training regimen. All training sessions were supervised by one of the researchers. Following five minutes of low-intensity treadmill running, subjects completed four sets of standing calf raise exercise, as described in the pre-testing section, with 30% of their pre-intervention 1-RM at full ROM to concentric muscle failure, either with or without BFR, according to their assigned group. Concentric muscle failure was defined as the inability to reach full ROM, which was controlled by a researcher during each set using a goniometer. In the BFR group, the cuffs were placed and closed before the first set, as previously described in the pre-testing section, and were not loosened or taken off during the rest periods until the fourth set was completed. Inter set rest periods for both groups were set at 30 seconds. Cadence of every repetition was set as 2-second eccentric and 2-second concentric, with no isometric hold at the top or the bottom of each repetition. This was monitored via mobile application Pro Metronome . The number of repetitions of all sets during each training session were documented in a training log. Pain was assessed after every set during the first session of the intervention using Visual Analogue Scale (VAS), as described by Heller et al. [All subjects had to attend at least of nine of the 12 (75% adherence) training sessions to be included for analysis. If participants missed two sessions in a row or more than three total, they were eliminated from the study. During the intervention, eight subjects missed more than three training sessions and were eliminated for further analysis . None of the subjects were injured during the RT sessions.Post-testing. Post-testing took place at least three days and not more than five days after the last training session. All procedures were identical to pre-testing. All researchers performing assessments were unaware of subjects’ intervention group allocation. Data from 21 subjects finishing the study were included for analysis. See Statistical analyses were performed using SPSS . Data were tested for normal distribution using Shapiro–Wilk test and homogeneity of variance using Levene’s test. Data are presented as means (SD). One-way analysis of variance (ANOVA) was used to test for baseline differences between groups. A 2×2 (time × group) ANOVA was used to determine differences in CV, MT, LS, and 1-RM. In addition, a 5x2 (time x group) repeated measures ANOVA was used to analyze the interaction of VAS values across time and group. An independent-samples t-test was conducted to compare the mean number of repetitions completed by the subjects per training session in both groups. A paired t-test was used to detect differences in the total number of repetitions in RT sessions 1 and 11. RT session 12 was not used for analysis as a number of subjects missed this session. Where necessary, Bonferroni post-hoc analysis was performed (adjusted for type I error). Within-group effect sizes (ES) were calculated as suggested by Dankel and Loenneke using the following formula: mean change score / SD of the mean change score . The chaIn addition, 95% confidence intervals (95%CI) for mean differences and mean percentage changes were calculated. Statistical significance was defined as p ≤ .05. Intraclass Correlation Coefficients (ICC) estimates were calculated based on a mean-rating (k = 2), absolute-agreement, 2-way random-effects model for MT and LS.Mean difference between the 50 repeated measures of MT across all participants, regardless of group assignment, was 0.001cm (0.05), and -0.64 kN/m (2.95) for LS. A high degree of measurement reliability was found. Mean measure ICC was 0.984 for MT, and 0.941 for LS respectively. Participants in the BFR attended a mean of 11.67 (0.49) of 12 possible trainings sessions and in the NoBFR 11.67 (0.50), respectively. Between RT session 1 and 11 the BFR group increased the total number of repetitions across all sets from 46 (7) to 59 (14) (p = 0.006), while in the NoBFR group there was no significant change ; RT session 2: 79 (19). The mean number of repetitions completed per training session was significantly higher in the NoBFR group (70 (16)) compared to the BFR group (52 (9)) (p = 0.002).There was no time x group interaction (p = 0.834), nor group effect (p = 0.447) in VAS. However, a time effect was observed (p<0.001) see . VAS in All other results are summarized in The BFR group improved their 1-RM from 163kg (37) to 203kg (42), and the NoBFR group from 163 (34) to 197kg (34). This equals a mean 1-RM increase of 40kg in the BFR , and 34kg in the NoBFR group respectively . Mean and individual changes can be found in The aim of this study was to investigate whether low-intensity RT with BFR is superior to RT without BFR to increase calf muscle strength and thickness in trained subjects. We could demonstrate no difference between training groups. In BFR and NoBFR, changes in 1-RM and MT were evident. This study demonstrates that the calf muscles are, in fact, trainable in terms of strength and hypertrophy.Significant increases in 1-RM were visible in both groups (BFR ES = 1.80 NoBFR ES = 2.50) of 24.80% in the BFR and 20.95% in the NoBFR group, respectively. There are studies showing a lower increases of knee extensor strength through comparable low-load BFR RT protocols (30% 1-RM) to failure. For example Clark et al.  showed aLow-intensity BFR and NoBFR resulted in changes of MT, even after only six weeks . The relative change mean change was 3.29% gain in BFR and 1.94% in NoBFR group. Other studies on hypertrophy following low-intensity BFR RT (20–30% 1-RM) of the quadriceps showed slightly higher results (between 6 and 7%) ,21,27. HCV remained unchanged in both groups. This is surprising, as significant changes in MT were present. It is also worth noting that CV changes were smaller than the usual measurement error of the device . This coTests of LS are a commonly used procedure to characterize stretch-shortening cycle (SSC) function , which pHowever, in contrast to Kubo et al. , more reVAS of pain increased significantly in both groups from baseline to the fourth set. This is in line with previous research . There mThe number of completed repetitions in a single session increased in BFR group over time (from 46 (7) to 59 (14)). It can be assumed, that through higher repetition volume, the principle of progressive overload was respected and lead to a sufficient training stimulus, even after six weeks. However, in the NoBFR group there was only a trend towards higher repetitions per session (71 (22) to 79 (19)). Nonetheless, adaptations were fairly identical in both groups. Whether differences would be present after longer intervention periods needs further research. The mean number of repetitions completed by the subjects per training session was significantly higher in the NoBFR group (70 (16) vs. 52 (9)), but in the usual range of this type of training . This isOne strong point of this study is the inclusion of RT-trained subjects only. This makes the results more applicable for populations with longer RT backgrounds. However, trained subjects have less potential for adaptation . NeverthIn conclusion, BFR and NoBFR RT lead to improvements of 1-RM and MT in RT trained males. LS and CV, however, remained unchanged. VAS values of pain in both groups were similar. For these reasons, it can be stated that BFR calf training is superior to NoBFR RT, as it leads to similar results but with less time, due to less repetitions required for completing a set until concentric muscle failure. Future studies should consider younger and older participants, trained and untrained, as well as women of all ages. In addition, researchers should consider using a time matched control group, longer intervention periods with different RT frequencies and training intensities. Further research on on RT periodization strategies, which include traditional high-intensity RT as well as low-load BFR RT on measures of strength, hypertrophy and athletic performance would also be a valuable contribution on the subject.S1 TableFour heartbeats were used to determine TA mean by Doppler ultrasound . Each condition was measured three times and the mean was calculated. The 7cm wide BFR cuff with a twist lock  was placed below the patella of the left leg and tightened to the maximum. The difference between “TA mean no cuff” and “TA mean with cuff” in percent equaled the resulting reduction in arterial blood flow velocity. BFR = blood flow restriction training; TA = Time averaged arterial blood flow velocity.(XLSX)Click here for additional data file.


It has been identified as one of the top 12 building-block chemicals. Here, we report the use of IA as a possible substitute to petroleum-based compatibilizers in polymer composite. We applied this study to thermoplastic elastomers based on styrene copolymers, since they are commonly used in blends and composites. Poly(styrene-b-ethylene-butylene-b-styrene) (SEBS) was grafted with 2.6 wt.% of itaconic acid (SEBS-g-IA) prepared by a reactive melt-mixing process, and was subsequently used to prepare composites filled with BaTiO3.). IA was successfully grafted as demonstrated by FTIR and XRD. SEBS-g-IA composites presented better mechanical properties, achieving an increase of Young modulus up to 80% compared with the neat polymer. This was ascribed to better dispersion and compatibility with the filler. Additionally, SEBS-g-IA showed increased dielectric permittivity, i.e., showed increased polarity, which indicates that it could potentially be used as a modifier for specialized polymers.Itaconic acid (IA) is an organic acid produced by the fermentation of sugars with  Most rd phase . SEBS isrd phase . Thus, ird phase .Aspergillus terreus and Aspergillus itaconicus. Consequently, itaconic acid is a renewable resource that could be useful for the synthesis of compatibilizing agents [3 nanoparticles.A commonly used and industrially available strategy to enhance affinity and compatibility is the inclusion of acid groups in the polymer structure. The most widely applied post-synthesis processing is the grafting reaction of maleic anhydride through reactive melt mixing ,6,7. Malg agents ,19,20,21g agents ,23. Here3) (<100 nm) were supplied by Sigma-Aldrich  and used as received from the suppliers.Styrene-ethylene-butylene-styrene block copolymer (SEBS) series Calprene H6110  was supplied by Dynasol Elastomers . Itaconic acid (IA (>98%), toluene, dicumyl peroxide (DCP) and barium titanate nanopowder  spectroscopy using a Perkin-Elmer Spectrum One spectrophotometer. The data were collected from 4000 to 700 cmλ = 1.5406 Å, with a power supply 40 kV and 40 mA. The incidence angle, θ, was fixed between 10° and 40° and the scan rate was 0.02°/s. The data were fitted using a Lorentz function in order to determine the solid mesophase contributions. The interplanar distance of solid mesophases was calculated by applying Bragg’s law:d was the interlayer distance and θ was the angle of the reflection plane. Likewise, the mesophase size, D, was determined using Debye–Scherrer equation:K was the shape factor (K = 0.9) and β was the line broadening at half the maximum intensity (FWHM).X-ray diffraction analysis was carried out using a Bruker D8 Advance diffractometer with a radiation source of CuK and wavelength Thermogravimetric (TGA) analysis was performed using a Netzsch thermogravimetric analyzer model Iris TG 209 F1. The temperature program was run from 25 to 800 °C at a heating rate of 5 °C/min in nitrogen atmosphere.−1 and a distance between clamps of 20 mm, according to the ASTM D412 specifications. Dog-bone-shaped specimens were cut from the membranes, having 35 mm length and 0.5 mm thickness. An average of five measurements for each sample was recorded.The tensile tests were carried out using an Instron dynamometer model 3366 at room temperature. The measurements were carried out at a crosshead speed of 200 mm/minThe morphology of the samples was studied using high resolution scanning electron microscopy, Inspect F50.−1 to 106 Hz at room temperature. Thin film disc-shaped samples were held in the dielectric cell between two parallel gold-plated electrodes. The amplitude of the AC electric signal applied was 1 V.Dielectric properties were performed on a broadband dielectric spectrometer model BDS-40, Novocontrol Technologies, GmbH, over a frequency range of 10The functionalization of SEBS with itaconic acid was carried out by a reactive melt-mixing process in the presence of dicumyl peroxide, which is a chain-growth polymerization initiator. The initiator provided free radicals, which reacted with abstracting tertiary hydrogen atoms present in SEBS’s backbone . Macrora−1, associated with the carbonyl stretching of the carboxylic acids of the itaconic acid [−1 associated with a five-membered anhydride ring [−1 associated with the C=C stretching of benzene ring of styrene moieties [nic acid ,26,27 anide ring . The graide ring . In addimoieties .cI), which is based on the Beer-Lambert law. For this, a calibration curve was built, by mixing different amounts of itaconic acid between 0.5 and 4.0 wt.% with SEBS by melt-mixing process. As shown in Equation (3), cI was estimated by considering the ratio between the integration areas of carbonyl absorption bands at 1770, 1743 and 1713 cm−1 and the C=C absorption band of styrene at 1601 cm−1 (−1) and carboxylic acid (1713 and 1743 cm−1) can be considered as the total percentage of itaconic acid grafted onto SEBS. Therefore, the percentage of itaconic acid grafted onto SEBS, IAG (wt.%), can be estimated using Equation (4), where the slope (1/0.1440) was determined experimentally by linear regression using the calibration curve displayed in The estimation of the percentage of grafting of itaconic acid onto SEBS was carried out by first establishing the so-called carbonyl index  was selected as compatibilizing after solubility tests were carried out in toluene and carbon tetrachloride. At this grafting content, SEBS-g-IA presented a good solubility compared to those with higher contents. It is known that SEBS is composed of hard and soft domains, where the hard segments are the polystyrene blocks and the soft segments correspond to the ethylene/butylene blocks. Both soft and hard segments form solid polymer mesophases. To study the influence of the grafting on the structure of solid mesophases, both samples were analyzed by X-ray diffraction . The peane phase ,30,31,323 in the presence of SEBS-g-IA as a compatibilizer. BaTiO3 behaved as a reinforcing agent, increasing the Young’s modulus and strength of SEBS and slightly decreasing its elongation at break. The addition of SEBS-g-IA induced a sensible increase of the mechanical properties of SEBS. This result was ascribed to the rigidity increase of the polar groups present in IA, which was also reflected in a decrease in the elongation at break of the material. Similar results have been observed by other authors when analyzing the mechanical properties of maleic anhydride grafted SEBS [3 composites. In the presence of the compatibilizer, Young’s modulus increases by almost 80%, probably because it favors the formation of interactions between its polar moieties, such as anhydride acid and carboxylic acid, and the surface of BaTiO3 particles. Such increase in the matrix polarity due to IA has previously been reported in an olefinic elastomer, in particular in ethylene-propylene copolymer [3 and SEBS/SEBS-g-IA/BaTiO3 present characteristic peaks at 22.2°, 31.6°, 38.9° and 45.3°, corresponding to (100), (110), (111) and (200) planes of cubic BaTiO3  [3 particles are coated with dopamine and dispersed in silicone rubber [3 particles. [BS-g-MA) . Other ae rubber . Additiorticles. .3 composites. SEBS-g-IA promoted the homogeneous dispersion of BaTiO3 in the SEBS matrix, which resulted in a sensible increase of the mechanical properties of the composite. Moreover, SEBS-g-IA increased the dielectric permittivity without significantly varying the dielectric loss. Therefore, SEBS-g-IA could be a promising route to improve compatibility and adhesion with other specialized polymers.SEBS successfully grafted with itaconic acid showed improved mechanical and dielectric properties and thermal stability. The grafting of itaconic acid affected the solid mesophases, giving rise to a polymer with more rigid domains. In addition, SEBS-g-IA behaved as an effective compatibilizer for the preparation of SEBS/BaTiO


Escherichia coli, in broilers kept under semi-field conditions, were examined. One-day-old broilers (Ross 308) (n = 400) were housed in four experimental rooms, subdivided in one seeder (S/C1)-pen and eight contact (C2)-pens. In two rooms, CE product was supplied from day 0 to 7. At day 5, seeder-broilers were inoculated with E. coli strain carrying blaCTX–M–1 on plasmid IncI1 (CTX-M-1-E. coli). Presence of CTX-M-1-E. coli was determined using cloacal swabs (day 5–21 daily) and cecal samples (day 21). Time until colonization and cecal excretion (log10 CFU/g) were analyzed using survival analysis and linear regression. Transmission coefficients within and between pens were estimated using maximum likelihood. The microbiota composition was assessed by 16S ribosomal RNA gene amplicon sequencing in cecal content of broilers on days 5 and 21. None of the CE broilers was CTX-M-1-E. coli positive. In contrast, in the untreated rooms 187/200 of the broilers were CTX-M-1-E. coli positive at day 21. Broilers in C2-pens were colonized later than seeder-broilers . The transmission coefficient between pens was lower than within pens . The alpha diversity of the cecal microbiota content was higher in CE broilers than in control broilers at days 5 and 21. The supply of a CE product from day 0 to 7 prevented colonization of CTX-M-1-E. coli after challenge at day 5, likely as a result of CE induced effects on the microbiota composition. Furthermore, compartmentalization reduced transmission rate between broilers. Therefore, a combination of compartmentalization and supply of a CE product may be a useful intervention to reduce transmission and prevent colonization of ESBL/pAmpC-producing bacteria in the broiler production pyramid.Extended spectrum beta-lactamase (ESBL)-producing bacteria are resistant to extended-spectrum cephalosporins and are common in broilers. Interventions are needed to reduce the prevalence of ESBL-producing bacteria in the broiler production pyramid. This study investigated two different interventions. The effect of a prolonged supply of competitive exclusion (CE) product and compartmentalization on colonization and transmission, after challenge with a low dose of ESBL-producing  Extended spectrum beta-lactamase and plasmid AmpC beta-lactamase (ESBL/pAmpC)-producing bacteria are resistant to extended-spectrum cephalosporins (ESC). ESBL/pAmpC-producing bacteria are present in humans, animals and the environment . PoultryEscherichia coli. This can be done by improving biosecurity. For example hygiene barriers can help reduce exposure to bacteria from the farm environment, or by cleaning and disinfection between production rounds. However, even after cleaning and disinfection, ESBL/pAmpC-producing bacteria might remain in the poultry house and result in colonization of the new flock , as well as transmission of ESBL-producing E. coli (E. coli (E. coli (Interventions can aim to reduce exposure of broilers to ESBL/pAmpC-producing ew flock . In addithe farm . Experimbroilers  and Stre in pigs . Furtherdditives  or compedditives . CE proddditives . In modedditives . Eggs ardditives . This su E. coli . Additio(E. coli . A prolo(E. coli .E. coli in young broiler chicks kept under semi-field circumstances. Two interventions were included: (1) prolonged supply of CE product from day of hatch until day 7, and (2) compartmentalization of a broiler flock. To investigate the effect of CE product on microbial composition, microbiota in cecal content was assessed before and after challenge by 16S ribosomal RNA (rRNA) gene amplicon sequencing.The aim of this study was to determine the effect of interventions on colonization and transmission of ESBL-producing Broilers were observed daily and the presence of clinical signs, abnormal behavior and mortality was recorded. The study protocol was approved by the Dutch Central Authority for Scientific Procedures on Animals and the Animal Experiments Committee of Utrecht University  under registration number AVD108002015314; all procedures were done in full compliance with Dutch legislation, and is thus compliant with legislation in the EU directive 2016/3/EU.n = 416), from a parent stock flock of 37 weeks of age, were transported directly after hatch to the animal facilities . Upon arrival, the broilers were individually tagged, weighed, and randomly divided over four experimental rooms (n = 104 broilers per room). Each room was subdivided into nine pens, with one seeder (S/C1)-pen in the middle , surrounded by eight contact (C2)-pens   broilers) were inoculated with CTX-M-1-E. coli and after 1 h moved back to the original S/C1-pens . Before the start of the experiment the parent flock, hatchery and research facilities were tested for the absence of ESBL/pAmpC-producing bacteria.Conventional broiler chicks  . The S/Cad libitum. The intervention was supplied in the drinking water (see section “Intervention Competitive Exclusion”); therefore, drinking water was not available ad libitum during the first seven days of the experiment in both intervention and control groups. Five out of 400 broilers died or were euthanized before the end of the experiment due to causes unrelated to the experiment, but by common causes in young broilers, i.e., not starting to eat or drink or omphalitis.Broilers were housed on fine wood shavings. A standard broiler diet without any antibiotics or coccidiostats, radiated with 9 Gy, was available ®, MSD Animal Health, Netherlands). From the moment of arrival in the rooms  until day 7, (4:00 p.m.), CE product was supplied in the drinking water, twice per day. Water solutions containing the CE product were prepared in pre-dilution, with a dose level according to recommendations of the manufacturer, i.e., 0.125 g CE product per 10 broilers. The amount of drinking water was restricted between day 0 and 7, based on the expected water consumption of 10 (C2-pen) and 20 (S/C1-pen) broilers in a pen to ensure that all supplied CE product would be consumed.In two of four rooms a CE product was supplied, containing “natural, live intestinal microflora derived from specific pathogen free (SPF) chickens and manufactured by fermentation” , isolated from conventional healthy broilers at slaughter age and resistant to cefotaxime  birds were exposed to 10 seeder birds, by moving the inoculated seeder birds to the corresponding S/C1-pens containing the contact birds.Broilers were challenged with fotaxime . Oral inpost mortem examination was done within 30 min after euthanasia for each broiler. Broilers were weighed and sex was determined, exterior and interior abnormalities were assessed, and ceca were collected and stored on dry ice for further analysis.Samples were taken using sterile dry cotton swabs . Broilers were sampled at day 5 at 4:00 a.m., just before inoculation to confirm absence of ESBL/pAmpC-producing bacteria, and from day 6 until day 21 daily at 8:00 a.m. At day 21, after the last sampling, n = 10) at day 5. At day 21, cecal content of all broilers in the S/C1-pen in all four rooms (n = 80) was collected. The closed side of one of the two ceca was cut and cecal content was gently squeezed into a 2 mL sterile cryotube and snap frozen on dry ice and stored at −80°C for genomic DNA extraction. To determine the microbial composition of the CE product, Aviguard® was suspended in PBS according to the manufacturer’s instructions and four aliquots of 2 mL were stored at −80°C for bacterial genomic DNA extraction. The full protocol for DNA extraction and determining microbiota composition was previously described , and the DNA samples were stored at −20°C until further use. Barcoded amplicons covering the variable regions V5–V6 and primers 784F and 1064R were used for 16S rRNA gene-based microbial composition profiling as previously described .All cloacal samples were enriched in 3 mL Luria-Bertani (LB) broth. After overnight incubation at 37°C, 10 μL broth were inoculated on MacConkey plates supplemented with 1 mg/L cefotaxime and incubated overnight at 37°C. n = 20) from the S/C1-pen and additionally 20 broilers from the C2-pens which were excreting CTX-M-1-E. coli. Samples were processed as previously described  and the ceca tested positive, broilers were assumed to be colonized at day 21. If only the ceca tested positive, broilers were not included as colonized birds within the time span of the experiment.Time until colonization was analyzed using parametric survival regression with an accelerated failure time model using a Weibull distribution . The hazDifferences of relative abundance were tested with Wilcoxon rank-sum test and corrected for multiple testing using Benjamini–Hochberg (BH) procedure. Alpha and beta diversity metrics were calculated and univariate and multivariate statistical analyses were applied to determine differences in the cecal microbiota. Alpha diversity (within sample richness) was determined using Faiths phylogenetic diversity, taking into account the phylogenetic relatedness . Differewithin and βbetween) were estimated based on a stochastic multi-pen SI model (The transmission coefficients for within and between pen transmission (βSI model  in whichpk) for a susceptible animal in pen k to become colonized during time interval Δt was calculated based on the force of infection (foi) within the pen and between pens (S/C1-pen to C2-pen):The probability  and in the adjacent pen connected with a mesh panel (cumexcrhoursadj) up to the beginning of the interval was used as a measure for environmental accumulation:In model 2 the within and βbetween in model 2 is 1/day2 and is interpreted as the number of new colonized broilers per day, caused by each day that one positive broiler has been excreting CTX-M-1-E. coli (The unit of β-E. coli .E. coli and CTX-M-1-E. coli (CFU/g) were tested using a linear regression model including the variables room, pen, sex, weight at day of hatch, weight at day 21, type of bird  and time until colonization. The best fitting model was obtained by backward selection based on lowest AIC value. The correlation between cecal content of CTX-M-1-E. coli and time until colonization was tested using Pearson’s correlation coefficient.The differences in cecal content of total E. coli. In the control groups all broilers in room 1 (n = 100), and 87/100 broilers in room 2 were colonized at the end of the experiment (Broilers in the CE groups (room 3 and 4) were not colonized with CTX-M-1-periment . Time unperiment .X2 = 1.90, p = 0.17), but the control broilers in room 1 had a lower alpha diversity than control broilers from room 2 . Within rooms, no differences between seeder and contact (C1) broilers were found. Weight at day of hatch, weight at day 21 and sex did not influence the alpha diversity (data not shown).The alpha diversity (phylogenetic diversity) was higher in cecal content samples of the broilers supplied with CE product (CE broilers) compared to the control broilers on day 5 and day 21 . On day R2 = 0.598, p = 0.009, uf: R2 = 0.688, p = 0.008). On day 21, application of the CE product explained 46% (wuf) and 51% (uf) of the variation between the cecal content samples . Within rooms, being a seeder or contact broiler did not explain any of the variation between the cecal content samples. The variation between the two control groups was larger than between the two intervention rooms (wuf: R2 = 0.351 vs. R2 = 0.210).In weighted and unweighted UniFrac (wuf and uf) distance based analysis, the supply of the CE product explained 60% (wuf) and 69% (uf) of the variation between the cecal content samples on day 5  were not colonized with CTX-M-1-between) was lower than the transmission coefficient within pens (βwithin) for both models. Model 2, with accumulated environmental transmission, was preferred over model 1, assuming direct transmission  and βwithin was 6.12 × 10–2 day–2 (95% CI 4.78 × 10–2 to 7.64 × 10–2)  .E. coli (log10 CFU/g) was lower in cecal samples from broilers from C2-pens than from the S/C1-pen, except for pen C26 and C27 (E. coli (log10 CFU/g) was lower in cecal samples from broilers kept in room 2 than broilers kept in room 1 . Broilers with a higher bodyweight at day of hatch had slightly higher cecal CTX-M-1-E. coli levels . Cecal CTX-M-1-E. coli levels were correlated with time until colonization, the shorter the time until colonization, the higher the cecal level . Mean E. coli levels in cecal content did not differ between rooms or pens.Mean CTX-M-1- and C27 . CTX-M-1E. coli after challenge of seeder birds. In the control group, 93.5% of the broilers were colonized at the end of the experiment. These results are in line with earlier experiments within isolators, in which a continuous supply of CE product during the first 14 days was able to prevent colonization  and reduced transmission, as previously shown in studies applying a single supply of CE  and the accumulative excretion time (model 2). The model including excretion time fitted better to the observed data, indicating that accumulation of ESBL-producing  chicken . In our E. coli. CE product could be supplied on the farm via the drinking water system. Supply should be done as soon as possible after hatching, before exposure to ESBL-producing E. coli occurs. In this study compartmentalization, including separation of feed- and water systems and strict hygiene measures, reduced transmission of ESBL-producing E. coli but could not prevent it. In practice, with less strict hygiene measures, the effects might be smaller.In poultry practice, the interventions studied in this experiment could be used to control the spread of ESBL-producing Overall, our study shows that CE is a useful intervention tool to prevent colonization of ESBL-producing bacteria after challenge with a low dose in the first week in a broiler flock. Transmission within a flock could be delayed by compartmentalization, however as soon as ESBL-producing bacteria are excreted and accumulate in the environment spread to other birds seems inevitable. Therefore, compartmentalization of large flocks into smaller groups of birds, which is for instance more common in breeding flocks at higher levels of the broiler production chain, could be combined to enhance the efficacy of other interventions. CE products could be supplied to young chicks after hatching at different levels of the broiler production pyramid to prevent colonization of birds. The insights provided by this study may provide a basis for further developments toward practically applicable measures to further reduce antimicrobial resistance in poultry.PRJNA647260.Raw sequence data were deposited into the Sequence Read Archive (SRA) at the NCBI, under accession number The animal study was reviewed and approved by Dutch Central Authority for Scientific Procedures on Animals and the Animal Experiments Committee of Utrecht University  under registration number AVD108002015314.AD, EF, JvdG, FV, DC, DM, and AS: experiment design. AD and FV: experiment execution. AD, JK, and EF: data analysis. AD and JK: manuscript writing. All authors discussed, read, contributed to, and approved the final manuscript.The views expressed are purely those of the author(s) and may not in any circumstances be regarded as stating an official position of the European Commission.DC is currently employed by the Research Executive Agency. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To compare the performance of an 18-gauge nonfenestrated catheter (18-NFC) with a 22-gauge fenestrated catheter (22-FC) for cardiac CT angiography (CCTA) in patients with suspected coronary heart disease.nd generation dual-source CT with arterial phase CCTA were included in this retrospective investigation to either an 18-NFC or 22-FC. In comparison to the 18-NFC, the 22-FC has three additional perforations for contrast agent dispersal proximal to the tip. We examined the two groups for differences in their average attenuation in the right and left ventricles  and in the atrium  as well as in the proximal right coronary artery (RCA) and the left main coronary artery (LM). The averages were calculated for both the 18-NFC and 22-FC.74 consecutive patients imaged on a 2Catheters were successfully placed on the first attempt 97% (36/37) for 18-NFC and 95% (35/37) for the 22-FC. The following enhancement levels were measured: 22-FC (in Hounsfield-Units (HU)): RV = 203±29, LV = 523±36, RA = 198±29, LA = 519±38, RCA = 547±26, LM = 562±25; 18-NFC: RV = 146±26, LV = 464±32, RA = 141±24, LA = 438±35, RCA = 501±23, LM = 523±23; RV , LV (p = 0.12), RA (p = 0.02), LA (p = 0.04), RCA (p = 0.3), LM (p = 0.33).No significant differences in attenuation levels as well as in image quality of the coronary arteries were found between NFC and FC. Nevertheless, the 22-gauge FC examinations showed significantly higher attenuation in the left and right atrium as well as the right ventricle. Patients with poor venous access may benefit from a smaller gauge catheter that can deliver sufficiently high flow rates for CCTA. Coronary CT angiography (CCTA) is an application that has traditionally required relatively high contrast media injection flow-rates –4. Thus,Therefore, the aim of this study is to compare the performance of an 18-gauge NFC with a 22-gauge FC for CCTA in patients suspected coronary artery disease. Retrospective data evaluation will be used as part of an objective image quality analysis to determine whether sufficient contrast can be achieved with CCTA using smaller-lumen catheters. This is of particular clinical interest when elderly patients with multi comorbidities need a diagnostic CCTA, while this cohort often shows limited peripheral venous conditions that do not allow the application of conventional large-lumen catheters.The study protocol of this retrospective investigation was approved by the local Institutional Review Board with a waiver of informed consent . The data was anonymized and retrospectively analyzed. Our study complied with the Declaration of Helsinki adopted by the World Medical Association (WMA) in 1964. A total of 74 patient CCTA examinations were analyzed in this study. The type of intravenous catheter used was identified via chart review along with data regarding catheter placement success and complications. Patients were included based on the following inclusion criteria: >18 years of age and CCTA performed between January 2016 and December 2017. Exclusion criteria were inadequate CCTA image quality, mainly caused by respiratory (n = 3) and cardiac motion artifacts artefacts (n = 2). These so-called "stair-steps-artefacts" can complicate sufficient density measurement at different locations and lead to incorrect measurement results.™ Pro Safety, Becton Dickinson) and a 22-gauge FC . In comparison to the NFC, the FC has three additional outlets for contrast agent dispersal just proximal to the terminal outlet. For the 22-gauge, each of the additional outlets measures approximately 0.76 mm in length and 0.25 mm in width. Two types of catheter were used: an 18-gauge NFC  at 120 kVp. The scan parameters were as follows: 120 kVp tube voltage, 50 mAs reference tube current using automated tube current modulation (effective mAs = 64 ± 31), 0.3 s rotation time, pitch 0.6, 128 x 0.6mm detector collimation. All images were reconstructed with a slice thickness of 1.5mm. Location of the catheter, CT-protocol, infusion rate, contrast volume and any catheter-related complications were recorded. Example complications include: exceeding maximum pressure, rupture of the perfusor extension line, contrast extravasation, or activation of the high-pressure alarm.Images were evaluated for both objective and subjective quality by a radiologist (8 years experiences in cardiovascular imaging). The aortic enhancement levels were measured by a CT-technologist using regions of interest (ROI) and controlled by a resident physician (2 years experiences in cardiovascular imaging). A ROI which covered 50% of the lumen was placed in the diaphragmatic section of the aorta. Additionally, the enhancement levels of both atria and ventricles were measured in the CCTA. For this purpose, the ROI was placed in the respective anatomical region in the axial sectional image plane in order to subsequently record the HU values .All statistical analyses were performed using dedicated software .Averages were calculated for both the standard 18-gauge NFC and 22-gauge FC. Categorical variables were expressed using frequencies and percentages.For both the 18-gauge NFC and the 22-gauge FC attenuation values were measured and the mean averages and standard deviations (SD) were calculated. A Student t-test was obtained to determine if there was a significant difference between the groups regarding their average attenuation.Comparison of infusion rate, contrast volume and exceedance of maximum pressure was performed using individual two-sample Student t-tests as well. A p-value of less than 0.05 was considered statistically significant.Patient examination of a total of 74 subjects 42 male) were included in this analysis. Between the 18-gauge NFC (n = 37) and the 22-gauge FC (n = 37) groups, there was no significant difference in age (p = 0.814), sex (p = 0.24), or contrast volume administered (p = 0.24)  was exceeded more often with the 22-gauge FC catheters ) than with the 18-gauge NFC ). This difference was statistically significant (p = 0.0281).Of note, there was an additional unexpected complication documented that occurred in the examination protocols: the extension line used, ruptured three times during contrast administration with 22-gauge fenestrated catheters . When this occurred, a new catheter was used to obtain intravenous access and the examination was attempted a second time with the same parameters. In all cases, the second examination attempt was successful . This failure rate was not statistically significant (p = 0.24).In all 74 CT-scans the subjective image quality was assessed and deemed acceptable. The aortic and cardiac enhancements levels could be measured for every subject .Regarding the mean attenuation, significantly higher values for the FC were found in the ROIs of the left atrium , right atrium  and right ventricle . No significant differences in attenuation were found for the ROIs of the left ventricle (p = 0.12), aorta (p = 0.28), right coronary artery (p = 0.3) as well as the left coronary artery (p = 0.3). All data of the comparison is shown in CCTA has become increasingly common in clinical practice as an essential diagnostic component –17. PartIn addition, there were no extravasations in either group. Some evidence suggests, as Wienbeck et al points to, an increased rate of extravasation in small-lumen 22-gauge catheters in the context of increased contrast medium flow . HoweverSeveral studies have shown a reduced risk of venous tear and consecutive extravasation in the presence of additional side-outlets, in contrast to the classic single-distal-opening catheter system, where the additional outlets allow flow rate and pressure to be distributed over several openings , 21. DanJohnson et al. examined contrasting of the aorta with an 18-gauge NFC and 20-gauge FC at an average infusion rate of 5.74 ml/s for the 18-gauge NFC and 5.58 ml/s for the 20-gauge FC . Both tyOur study underlies various limitations. First, catheter types were compared between different patients, but not individually in every patient. This is mainly due to the fact, that most patients did not receive multiple CCTAs at our medical center. Ideally, every patient would have received two scans: once with an NFC and once with a FC. Differences in canula placement, enhancement and image quality might therefore be biased by overall group differences. Regarding the study design, a completely blind evaluation of catheter placement was not possible as there was an obvious visual difference between the two catheter types. However, the image evaluation was performed blinded. Furthermore, the increased pressure load of the 22-gauge FC in the study resulted in minimal, temporal delay of the contrast agent peak, even though we tried to maximize the potential pressure load through the bolus tracking technique. Another factor in the use of FC is the cost factor, which is many times higher than a conventional catheter. Since this is a retrospective data evaluation, we cannot make any statements about the duration of the application of the respective catheter systems, as this was not recorded. In clinical practice, however, there is no significant time difference with regard to the duration of application. Future evaluations should take the higher cost factor and potential time savings in catheter placement into account. Regarding our sample size, the small number of 37 patients in each group limits the statistical power of the study. Although the antecubital region is the preferred location for catheter placement, the exact positions could not be deduced retrospectively from our reports. As the position of the indwelling venous cannula has an influence on the contrast, thisshould not be omitted as a further limitation of our study. As not the same person was responsible to establish all 78 i.v. catheters, different grades of expertise might have influenced the success rate at the first attempt.In summary, we were able to show that 22-gauge FCs are equivalent, and partially better in terms of contrast-enhanced image quality and safety compared to a conventional 18-gauge NFC at infusion rates of 5 ml/s in context ofCCTA. The administration of contrast agent with FC may better withstand the recognizable trend of increasingly faster acquisition times with MDCT scanners. With progressively aging patient cohorts, the rise of cardiovascular comorbidities and thus limited peripheral venous conditions, the use of FC might lead not only to faster but also more tolerable examinations due to reduction of catheter sizes. The evaluation of these long-term benefits in clinical praxis could be akey approach for further prospective studies.


Characterizations by chronoamperometry, UV-Vis spectroscopy and Raman spectroscopy showed that polypyrrole is stable under anodic and cathodic conditions, but CoTPPS and TPPS immobilized in PPy are degraded during the anodic process. Thus, decreases in photocurrent of up to 87% and 97% for CoTPPS-doped PPy and TPPS-doped PPy were observed after a 30-min chronoamperometry test. On the other hand, good stability of CoTPPS and TPPS immobilized in PPy was observed during photoelectrochemical oxygen reduction, which was reflected in almost constant photocurrents obtained by chronoamperometry. These findings are relevant to understanding the role of CoTPPS as a catalyst or pre-catalyst in photoelectrochemical applications such as water splitting. In addition, these results could pave the way for further research to include CoTPPS-doped PPy in the design of novel photocathodes.Cobalt porphyrins have emerged as promising catalysts for electrochemical and photoelectrochemical applications because of their good performance, low cost and the abundance of cobalt in the earth. Herein, a negatively charged porphyrin meso-tetra-(4-sulfonatophenyl)-porphin (TPPS) was immobilized in polypyrrole (PPy) during the electro-polymerization, and then it was metallized with cobalt to obtain meso-tetra-(4-sulfonatophenyl)-porphyrinato cobalt (II) (CoTPPS) as a dopant in PPy. The coatings were evaluated as photoelectrodes towards thiosulfate oxidation and oxygen reduction. For comparison purposes, the photoelectrochemical behavior of ClO In both cases, photoelectrochemistry has emerged as an attractive alternative that makes use of solar radiation to boost the electrochemical process ,2,3. In  and CdS ,5. With  and CdS ,7,8.The search for better performances in electrochemical and photoelectrochemical processes has led to the evaluation of alternative compounds such as porphyrins ,10. When3/TiO2. In this heterojunction the efficiency of hole injection in the electrolyte increased 41% compared with the heterojunction without the porphyrin. The authors reported the loss of the signals of the porphyrin (observed by UV-Vis spectroscopy) after 1 h of photolysis. This was attributed to the catalyst desorption from the photoanode [4/Al2O3 in a Co-meso-tetra(4-carboxyphenyl)porphyrin (CoTCPP) solution. In this case, a two-fold enhancement in photocurrent at 1.23 V vs. RHE was achieved in comparison with the yield of BiVO4/Al2O3 [P. Wei et al. reported the use of Ni-meso-tetra(4-carboxyphenyl)porphyrin (NiTCPP) as a water oxidation catalyst (WOC) incorporated by soaking on the system WOotoanode . SimilarO4/Al2O3 .X catalyst due to the degradation of three different cobalt porphyrins deposited on fluorine-doped tin oxide (FTO) [Although there are relatively few reports of metal porphyrins in photoelectrochemical water splitting, a higher amount of papers have addressed the water oxidation process with porphyrins without the use of light ,14,15,16de (FTO) , and T. de (FTO) .2 nanotubes with 5-(4-carboxy-phenyl)-10,15,20-triphenylporphyrinato copper(II) (CuTPP–COOH) and they achieved a photocurrent of 9 μA/cm2 at −0.3 V vs. NHE using only visible light (λ > 395 nm) [2 (at 0.5 V vs. RHE) was achieved under AM 1.5 and 1 sun illumination. Since no photocurrent was observed for bare NiO, the photocurrent achieved was attributed to the TCPP porphyrin and the match between the reduction potential of TCPP and the valence band of NiO [In photoelectrochemical oxygen reduction, some studies have shown the potential of porphyrins to carry out the reaction. In these systems (as in water splitting) porphyrins with carboxyl groups are frequently used to improve the attachment of the porphyrins to the electrode surfaces. D. H. Apaydin et al. coated TiOd of NiO .Although porphyrins substituted with carboxyl groups have shown the best photoelectrochemical performances, it has been observed that the interactions between the semiconductor surface and the carboxyl groups are not strong enough to ensure good attachment. In this way, alternative strategies such as the entrapment of charged porphyrins in conductive polymers are very attractive for the development of novel photoelectrodes.4−-doped polypyrrole films were also synthesized. Since porphyrins have gained attention in photoelectrochemistry, we evaluated for the first time the photoelectrochemical stabilities of CoTPPS and TPPS entrapped in PPy under anodic and cathodic conditions to perform thiosulfate oxidation and oxygen reduction, respectively. Polypyrrole was selected because of its low cost, good processability, ease of synthesis and good performance as catalyst support [Herein, a negatively charged porphyrin meso-tetra-(4-sulfonatophenyl)-porphin (TPPS) was immobilized in polypyrrole (PPy) during the electro-polymerization process, then it was metallized with cobalt to obtain meso-tetra-(4-sulfonatophenyl)-porphyrinato cobalt (II) (CoTPPS) as a dopant in PPy. For comparison purposes, ClO support ,23,24. Tw/w NaOH  solution at 55 °C, after that, the substrates were sonicated twice in ultrapure water for 15 min. Pyrrole  was distilled and stored under a nitrogen atmosphere. Acetonitrile  was kept over molecular sieves. LiClO4  and (CH3COO)2Co.4H2O  were used without further purification.FTO substrates 25 mm × 12.5 mm  δ 8.86 , 8.20 , 8.06 , −2.94   δ 147.79, 141.33, 133.77, 124.24, 119.74 : 414, 516, 553, 581, 635  . 13C-NMR, 119.74 . UV-Vis 581, 635 .2) using a precursor solution composed of 0.1 M pyrrole and 0.1 mM TPPS in deionized water. For ClO4−-doped PPy the electrosynthesis was accomplished under 0.5 mA/cm2 using a solution composed of 0.25 M pyrrole and 0.5 M LiClO4 in acetonitrile +2% w/w H2O. For the obtention of CoTPPS-doped PPy a previously reported route was used [2+ ions were introduced into the porphyrin by immersion (10 min) of TPPS-doped PPy in aqueous 0.1 M cobalt acetate solution heated to 90 °C.Pyrrole electro-polymerizations were carried out in a conventional three-electrode cell by using FTO substrates as working electrodes (WE), a platinum foil as counter-electrode (CE) and an Ag/AgCl electrode as reference electrode (RE) . The thiwas used . In this2S2O3 and 0.1 M Na2SO4 for anodic and cathodic polarizations, respectively. For illumination (100 mW/cm2), a solar simulator ABET technologies 10500  was used. UV-Vis spectra were measured with an Analytik Jena SPECORD 50 PLUS spectrophotometer . Raman spectra were taken using a 532 nm laser in a HORIBA Scientific XploRA equipment . IR spectra were obtained with an IRTracer-100 FTIR Shimadzu spectrophotometer .The morphology of PPy coatings was examined by scanning electron microscopy (SEM) using TESCAN LYRA3 equipment . Photoelectrochemical characterizations were performed in 0.1 M Na4−-doped polypyrrole (PPy-ClO4) and TPPS-doped polypyrrole (PPy-L) were carried out in both cases supplying 42, 31 and 21 mC/cm2. Since the thickness of the coatings is directly related to the electric charge supplied, thicknesses of 87 nm, 64 nm and 43 nm were calculated, respectively [The electrochemical synthesis of ClOectively .4, suggesting more favorable conditions for electro-polymerization.The galvanostatic conditions that were used produced homogeneous polypyrrole coatings on FTO substrates. Moreover, the potentials reached during the growth of the films were low enough to ensure that polypyrrole over-oxidation did not occur . This po−1 wavenumbers, but these signals disappeared after metalation and a new absorption band was appreciated in 957 cm−1. This metal-dependent band has been previously assigned for other porphyrins as an in-plane porphyrin deformation mode [After the synthesis of PPy-L, the insertion of cobalt ions in the porphyrin was carried out to obtain CoTPPS-doped polypyrrole (PPy-C). This process did not lead to visible changes in the coatings. However, the IR spectra of the composite material showed some changes with the incorporation of cobalt . In the ion mode ,29. As a4 presented a structured shape with many and small globules on the surface (globular topology), although the shape of the FTO grains was still identified. In the case of PPy-C and PPy-L, both had a morphology with a planar tendency in which the coatings followed the shape of the FTO surface. In all cases, no uncovered areas of FTO were observed.On the other hand, the morphology of the films was examined by scanning electron microscopy . PPy-ClO2. It can be observed that the bare FTO presented current densities near zero during the potential sweep. In contrast, all the coatings showed responses with and without illumination. Based on this, it can be suggested that these polymeric films have catalytic activity towards thiosulfate oxidation, while their photocatalytic properties were evidenced in higher current densities when irradiation was used.The photoelectrochemical performance of the composite materials under anodic conditions was examined by linear sweep voltammetry (LSV) and chronoamperometry measurements. 4. This trend was also observed in the coatings obtained with 31 mC/cm2  when an electric charge of 42 mC/cm2 was used. However, it decreased rapidly as the thickness of the coating was reduced, so that ~4 μA/cm2 was obtained using 21 mC/cm2. On the other hand, PPy-C had a slower loss of stability than PPy-L as shown by the behavior of photocurrents over time. This indicates that the presence of the cobalt complex changes the photoelectrochemical properties of the PPy coatings. However, if the thickness of the coating is related to the amount of complex entrapped, it follows that this amount does not noticeably influence the photoresponse of the electrode. In the case of the dark currents observed during all the characterizations by chronoamperometry, they can be attributed to the catalytic activity of the films to carry out the reaction. However, in the case of PPy-L and PPy-C, the dark currents could be associated with their degradation process.The stability and magnitude of the photocurrents according to the dopant and thickness of the films was evaluated by chronoamperometry (at 0.6 V vs. Ag/AgCl) using chopped light. For this purpose, six changes from darkness to illumination were distributed during the characterization as can be seen in  coating D. This w coating E and PPy coating F since a4 to have higher currents densities by LSV, given its globular topology that confers a high surface area . For this purpose, photoelectrodes of PPy-ClO4, PPy-L and PPy-C were synthesized with 42 mC/cm2. Characterizations were carried out in solutions previously bubbled with N2 and with air for 15 min to analyze the activity of the photoelectrodes towards oxygen reduction . Our results show the potential of these composite coatings in the electrocatalysis of light-assisted cathodic reactions. They also suggest that these materials are not suitable in anodic reactions, such as photoelectrochemical oxidation of water. In the case of ClO4−-doped PPy, dark currents and photocurrents showed stability under anodic and cathodic conditions, which reinforces the strength of polypyrrole as a matrix material in the development of photoelectrodes.The cobalt porphyrin CoTPPS and its free-base porphyrin TPPS were entrapped in polypyrrole by electropolymerization. This allowed obtaining CoTPPS-doped PPy and TPPS-doped PPy with controlled thicknesses. Photoelectrochemical and spectroscopic characterizations showed that CoTPPS and TPPS in PPy degrade easily under anodic conditions, which was reflected in the poor stability of the photocurrents during oxidation of the electron donor thiosulfate . On the other hand, it was found that CoTPPS and TPPS entrapped in PPy are stable in photoelectrochemical oxygen reduction since nearly constant photocurrents were obtained by chronoamperometry (16 μA/cm


In experiments on cardiac fibers treated with the hypertrophic cardiomyopathy-related mutants E244D- and K247R-troponin-T, treatment with K247R-troponin-T did not recruit contracting actomyosin to a greater extent than wild-type-troponin-T, although a similar drop in the intensity of the troponin reflection occurred. Therefore, the conformational change in K247R-troponin-T was suggested to be unable to fully recruit actomyosin interaction, which may be the cause of cardiomyopathy.X-ray diffraction and tension measurement experiments were conducted on rat left ventricular skinned fibers with or without “troponin-T treatment,” which exchanges the endogenous troponin T/I/C complex with exogenous troponin-T. These experiments were performed to observe the structural changes in troponin-T within a fiber elicited by contractile crossbridge formation and investigate the abnormality of hypertrophic cardiomyopathy-related troponin-T mutants. The intensity of the troponin reflection at 1/38.5 nm Deciphering the regulatory mechanism from a structural aspect is important for understanding the pathology of striated muscle diseases related to mutations of regulatory proteins, including familial cardiomyopathy. As for troponin, which is involved in the pathogenesis of familial cardiomyopathy, pieces of structural evidence that suggest formation of contractile crossbridges is involved in the regulatory mechanism of contraction have been accumulated to date ,3,4,5,6.2+, since the inhibitory effect of troponin I (TnI), which can be released by Ca2+ binding to TnC under physiological conditions, is now removed by the exchange. First, we confirmed that the active actomyosin interaction caused by addition of ATP in TnT-treated fibers elicited a structural change in troponin-T in a cooperative manner, as has been reported with skeletal muscle [In this study, we performed X-ray diffraction experiments on carefully prepared cardiac skinned fibers devised to obtain detailed X-ray diffraction patterns. Furthermore, we applied the method of “troponin-T (TnT) treatment”  to thesel muscle ,13. Secol muscle  whereas l muscle ,16. Equa−1 on the meridional axis, troponin reflection was observed . In Diffraction patterns of cardiac muscle fibers with no treatment before and after addition of ATP are shown in Next, we examined the diffraction patterns from TnT-treated cardiac muscle fibers. Diffraction patterns obtained from cardiac fibers treated with wild-type TnT before and after the addition of ATP are shown in Finally, we examined the changes in the diffraction pattern of cardiac fibers treated with E244D-TnT and K247R-TnT mutants to obtain information about the pathological mechanism of HCM-related TnT mutations. The intensity profile of the troponin reflection along the equatorial axis and integrated intensity of TnT-treated cardiac fibers with E244D-TnT and K247R-TnT are shown in To evaluate the number of recruited crossbridges in the fibers after the TnT-treatment, the tension generating capacity and intensity ratio of the 1.1 and 1.0 equatorial reflections were analyzed. As seen in the representative tension trace after the TnT treatment in Significant intensity change of troponin reflection was induced by addition of ATP in cardiac muscle fibers after TnT-treatment, indicating that troponin-T undergoes a structural change triggered by formation of contractile crossbridges in the cardiac skinned fibers. It is known that regulation by the tropomyosin-troponin system is influenced by formation of active crossbridges ,13, implAs for the intensity drop, Matsuo et al. , from thRegarding this study, the intensity change of the 38.5 nm troponin reflection observed with TnT-treated fibers was considered to be due to a structural change of TnT because the remaining endogenous T/I/C complex is unlikely to cause a large intensity drop A,E. HoweThe relative 1.1/1.0 intensity ratio and tension measurement suggested that cardiac fibers treated with K247R-TnT exhibited a weaker property of contractile crossbridge recruitment, contrary to the anticipation based on reports by Matsumoto et al.  and FordNext arises the question of how this weaker property of contractile crossbridge recruitment causes K247R-TnT to induce cardiomyopathy. It is notable that K247R-TnT was considered as an HCM-causing mutant based on the first report of this mutation in HCM-patients by Garcia-Castro et al. ; howeverThe results of the 1.1/1.0 intensity ratio and tension development experiments imply that introduction of E244D-TnT would confer the trend to elicit larger crossbridge recruitment than the wild-type TnT B,D. The Matsuo et al. , based o−1), following which their hearts were rapidly excised and blood was discharged by beating in warmed and oxygenated artificial extracellular solution. Next, cardiac muscle fiber bundles were dissected from the left ventricle wall very carefully. Cell membrane was disrupted by treatment of the fibers for more than 3 h with artificial intracellular solution containing 0.5% triton-X 100.Male rats  were used with approval of the Institutional Animal Care and Use Committee of The Jikei University . The rats were deeply anesthetized by intraperitoneal injection of pentobarbiturates  for an average period of 5 h . Recombi2+, with ionic strength of 0.2 at pH 7.0, and in the ATP solution, which contained 3.5 mM MgATP and was nominally free of Ca2+,with ionic strength of 0.2 at pH 7.0, using an imaging plate system . The X-ray wavelength was 0.09 nm and the beam size was 250 µm in width and 150 µm in height. The specimen-to-detector distance was 1.8 m. The sarcomere length of each fiber bundle was adjusted to 2.3 µm on average by laser diffraction. Experiments were carried out at 20 °C.X-ray diffraction experiments were performed at BL45XU in SPring8  using a setup described elsewhere  with sli−4.4 M Ca2+, with ionic strength of 0.2 at pH 7.0, was used.Tension development in the cardiac muscle skinned fibers with or without TnT-treatment was measured using a tension transducer . To evaluate maximally activated tension of the fibers, Maximally-activating solution, containing 3.5 mM MgATP and 10−1 along the meridional axis was integrated within a slice of 0.00081 nm−1 width in the equatorial direction. Then, two Gaussian distributions along the meridional axis were fitted to the profile to obtain the troponin-derived reflection separately from that of the C-protein-derived reflection for each slice. This was repeated after shifting the slice by 0.00081 nm−1 in the equatorial direction.Diffraction patterns were analyzed using MATLAB R2016B. Each X-ray diffraction pattern was rotated around the center and averaged quadrantally. The intensity profiles of the 38.5 nm troponin reflection along the meridional axis were obtained as below. Intensity in 1/54.8–1/32.9 nm−1.The obtained intensity of the troponin reflection for each slice was plotted along the equatorial axis in the range of 0–1/47.4 nm−1 along the meridional axis was integrated within a slice of 0.010 nm−1 width in the equatorial direction and the peak area above the background line between the bottoms at either side of the peak was obtained for each slice. This procedure was repeated after shifting the slice by 0.010 nm−1 in the equatorial direction. The obtained intensity for each slice was integrated along the equatorial axis in the range of 0–1/4.9 nm−1. Intensity of the 38.5 nm meridional reflection of troponin was expressed relative to the total integrated intensity of the 5.9 nm actin layer line.The intensity profile of the 5.9 nm actin layer line was obtained as below. Intensity in 1/5.6–1/6.3 nm−1 in the meridional direction. Next, intensities of the 1.0 and 1.1 equatorial reflections were obtained by fitting two Gaussian distributions on an exponential background to the integrated profile.Equatorial intensity profiles were obtained by integrating intensities in the range of 0–1/61.7 nmThrough direct observation of structural changes in TnT within the cardiac fibers, the K247R-TnT mutation was suggested to have a lower recruitment capacity of contractile crossbridges, even though a similar level of intensity drop in the troponin reflection occurred. Thus, the alteration of the single residue, K247R, affected function of the tropomyosin-troponin regulatory system without a large conformational change in TnT.2+ sensitivity, is beyond the scope of this study. Future work on cardiac muscle fibers with mutant TnT and normal troponin I/C will further clarify the molecular pathology of TnT-related cardiomyopathy.Finally, it should be noted that this study is limited to the function of TnT. Its interaction with troponin I/C, which affects actomyosin kinetics and Ca


Bundesliga season. The levels of creatine kinase (CK), urea, and C-reactive protein (CRP) were measured during 3-wkmid and 3-wkend each Monday, Wednesday, and Friday. In 3-wkmid the CK median was 14% higher (241 vs. 212 U/L) compared to 3-wkend . Overall, the medians of CK, urea , and CRP  during 3-wkmid did not differ to the values of 3-wkend. Daily coefficient of variations (CVs) ranged from 22 to 71% (CK), 17 to 37% (urea), and 9 to 164% (CRP). Individual medians ranged from 101 to 350 U/L (CK), 23 to 50 mg/dL (urea), and 0.6 to 1.1 mg/L (CRP). High intra-individual variability was demonstrated by large intra-individual CVs . Our data show (i) large inter-day and inter-week variability of all biomarkers, depending on the external load and (ii) considerable inter- and intra-individual parameter variations. Creatine kinase concentrations could sensitively reflect soccer-specific loads during the season.The aim of this study was to investigate inter-day and -week as well as intra- and inter-individual variation of selected biomarkers in high-performance youth soccer players to assist practitioners interpreting player’s internal load to counteract underperformance and unwanted health risks. Eleven male youth soccer players were tested multiple times during two 3-week periods at midpoint (3-wk Soccer players under the age of 19 (U-19) are exposed to frequent training sessions and matches, which are scheduled in one- or two-match weeks. It is well known that high match and frequent training session exposure in youth soccer players is associated with increased injury risks .Monitoring internal and adjusting external load may ensure optimal training and match performances as well as prevent injuries over the course of an entire season. In this regard current expert opinion warrants objective and reliable methods (i) to adjust external load for optimal adaptation and prevention of overuse and (ii) to quantify each player’s fatigue state . From a Previous studies with professional soccer players revealed acutely increased creatine kinase (CK), urea, and C-reactive protein (CRP) concentrations induced by soccer-specific loads  and by vTherefore, in-season monitoring of CK, urea, and CRP may assist estimating the internal load of muscles, metabolism, and unspecific inflammatory condition . AdvanceThis exploratory study aimed to compare two selected time periods within a national U-19 soccer season to investigate (i) the inter- and intra-individual kinetics of selected biomarkers and (ii) the seasonal inter-day and inter-week variability of CK, urea, and CRP. From a practical perspective the study results should assist assessing high-performance youth soccer players’ internal load to counteract underperformance and unwanted health risks.Eleven male youth soccer players of a German U-19 first league (1. Bundesliga) team were tested  during two 3-wk periods. Two players were members of the U-19 national teams of Germany and Poland. We investigated only field players since physical requirements vary widely between field players and goalkeepers. All players were informed about all testing and signed an informed consent. This study was approved by the ethics committee of the German Sport University  and was conducted according to the Declaration of Helsinki.mid) and at the end of the first half of the season (3-wkend) during a German U-19 first league season. A total of 3-wkmid took place between match 5 and 8, 3-wkend between match 10 and 14, which was at the end of the first half season . Two first league matches were scheduled before the 3-wkend (rounds 10 and 11 were played on Thursday 6.30 p.m. and Sunday 1.00 p.m.).Before and after each week of 3-wkBecause of the observational nature of this study, data were analyzed retrospectively. Therefore, the present results had no immediate consequences for coaches and medical staff to adjust training load, medication or recovery strategies by coaches or the medical staff.mid and 3-wkend on every Monday (Mon), Wednesday (Wed), and Friday (Fri) capillary blood was sampled from the participants’ earlobe before the beginning of each training session and analyzed immediately after sampling. Training sessions commenced at 5.00 p.m on Mon, at 11.30 a.m. on Wed, and at 3.00 p.m. on Fri. Only on Wed1  the training began at 5.30 p.m. On Wed6  all players had one day off, so all testing was canceled by the coaches.During 3-wk® Plus system . The measuring ranges were 24.4 to 1,700 U/L (CK) and 20.0 to 300.0 mg/dL (urea). The intra-assay coefficient of variation (CV) (test-retest reliability) for CK was 3.5% and for urea ≤3.0%. The inter-assay CV (day-to-day reliability) for CK was ≤4.2% and for urea ≤3.5%. QuikRead go® hsCRP + Hb tests for a QuikRead go® photometer  determined CRP concentration from 20 μL capillary blood sample. During the last two weeks of the study QuikRead go® wrCRP + Hb tests were used, whereby only 10 μL of blood were needed. Measuring ranges were 0.50 to 75.00 mg/L (hsCRP), and 0.50 to 300.00 mg/L (wrCRP), respectively. The intra-assay CV for hsCRP ≤ 5.9% and for wrCRP ≤ 4.9%. The inter-assay CV for hsCRP ≤ 2.7% and for wrCRP ≤ 4.8%.Creatine kinase and urea concentrations in each 32 μL blood samples were analyzed employing ReflotronP = 0.001 and P < 0.001, respectively), CRP , and urea  concentrations. Therefore, all data are presented in box plots as medians, 25 to 75% percentiles as well as minimum and maximum values. Inter-day and inter-week variability are demonstrated by boxplots which represent the results from every testing day. Intra- and inter-individual variability are demonstrated by boxplots representing the results from every player during the observation period. The CV quantified the parameter variability and was defined as standard deviation divided by mean value multiplied by 100. CV was determined for all biomarkers during the complete study process as well as for 3-wkmid and 3-wkend and for all testing days. CV was calculated from the individual parameter responses in the study process to quantify intra-individual variability. The differences between 3-wkmid and 3-wkend were assessed employing Wilcoxon comparison. The effect size (ES) measurement for non-parametric tests (r = Z/√N)  and Excel 2010 , respectively.Data distribution with Kolmogorov–Smirnov and the Shapiro–Wilk test showed no normal distribution of CK ( = Z/√N)  were calmid and 3-wkend).mid and 3-wkend. Overall, the medians of CK , urea , and CRP  in 3-wkmid did not differ compared to the values of 3-wkend.mid and 3-wkend .The external load did not differ between 3-wkmid) to 147% .During both observation periods the biomarkers ranged from 47 to 791 U/L (CK), 20 to 71 mg/dL (urea), and 0.5 to 11.0 mg/L (CRP), respectively. CVs ranged from 33 , Wed1: 264 U/L (+ 59%), and Fri1: 166 U/L). CRP medians in week 1 and 2 were also higher on Mon and Wed compared to Fri , Wed2: 0.9 mg/L (+ 42%), and Fri2: 0.6 mg/L). In 3-wkmid daily mean CK and CRP values showed very large correlation . In 3-wkend daily mean CK and CRP values showed large correlation but not significant .end urea medians on Fri4  and Fri5  were higher than on Wed4 (31 mg/dL) and Wed5 (36 mg/dL), respectively. Daily CVs ranged from 22% (Fri5) to 71% (Mon4) for CK, from 17% (Fri6) to 37% (Mon1) for urea and from 9 (Wed4) to 164% (Fri5) for CRP concentrations.In 3-wkOne day after the matches (Mon2 and Mon3) median CK values (337 and 299 U/L) were about 70% higher than 2 days postmatch  with CK medians ranging from 152 to 229 U/L.One day after match day median CRP values (1.2 and 0.8 mg/L) were about 43% higher than after 2 days (range from 0.5 to 1.0 mg/L), while median urea values were nearly unchanged between one (36 and 34 mg/dL) and 2 days (range from 30 to 41 mg/dL) postmatch.The individual kinetics in Individual CVs in To illustrate inter- and intra-individual variability, only players with at least 11 analyzed blood tests were included.The aim of the present study was to compare the results of two selected time periods within a soccer season, to investigate the inter-day and inter-week variability of selected biomarkers  and to investigate the inter- and intra-individual kinetics of each variable. The main findings of this study were: (i) large inter-day and inter-week variability of all biomarkers and (ii) considerable inter- and intra-individual parameter variations.Here we measured large inter-day variability of CK, urea, and CRP concentrations in youth players during two competition 3-wk periods. Based on the large variability we conclude that testing of CK, urea, and CRP should be performed as frequently as possible to (i) understand the kinetics of each variable during a season and (ii) generate meaningful consequences for training and match tactics. For example, one blood test per week may assess the current status of CK, urea, and CRP, but single sampling prevents to judge whether the concentrations are increasing, peaking or decreasing. Therefore, when applying single measurements, e.g., one test each week, the potential for data interpretation – and consequently for practical consequences – is severely limited  and Sunday (one day postmatch) impacts CK responses on Mondays at the beginning of the training week, especially for the players with more than 45 min match participation. As seen in Overall, it seems that youth soccer players need approximately 24–48 h to recover after an official  or simulend (Fri5: 43 mg/dL) was reached after three successive training days. As seen in Urea inter-day variability is affected by load intensity and duration . FurtherThe CRP inter-day variability shown in this study can be explained by the differing load stimuli  in the dmid reflected a certain periodization model planned by the coach’s external load through increasing loads in three successive weeks. This observation is supported by elevated median values 3 weeks in a row based on the weekly average, and by the Wednesday results, showing that soccer-specific loads are sensitively reflected by the players’ CK levels when frequent and dense measures are performed. In contrast, no periodical CK variations were shown in the last three testing weeks. This could be argued with the fact, that the weekly training plans in 3-wkend showed an altered structure compared to 3-wkmid. For instance, in 3-wkend the days with no training sessions varied from week to week .Considerable differences between CK, urea, and CRP responses were found in successive testing weeks . To the best of our knowledge no other publication U-19 soccer players has examined the inter-week variability of selected biomarkers by frequent blood sampling. In the present study, periodical CK variations were shown in the first three testing weeks, which are most probably a result of the varying external loads as seen in mid and 3-wkend between the mean daily CK and CRP values , which can be explained by the fact that muscle damage is often accompanied by local inflammatory responses , 13% (urea), and 29% (CRP), respectively. Because of the practical advantages of this method , POCT seems to be a practical method to monitor CK, urea, and CRP to judge high-performance U-19 players’ individual physical stress responses. Internal biomarker analyses provide evidence for the recovery status of biological systems, which cannot be analyzed by other external monitoring procedures . Consequently, in accordance with others , we recoBecause CK, urea, and CRP showed a large inter-day variability we advise the assessment of all variables as frequently and densely as possible in order to identify trends in their kinetics. For this reason, our recommendation is not to assess the biomarkers within a (single) status diagnostic procedure but rather by frequent in-season monitoring. CK, urea, and CRP concentrations should be interpreted by intra-individual kinetics and not by general sport-specific cut-off references. The CK concentrations seem to respond sensitively to soccer-specific loads. CK concentrations offer interesting and additional information which can be utilized to optimize the soccer player’s individual internal load, which may allow the prevention of fatigue-induced underperformances and injury.Because CRP detectability in blood is shorter compared to that of CK , and because CRP seems to be less sensitive than CK in reflecting external loading, our recommendation is not to frequently assess CRP levels. The possibility to assess CRP promptly and effortlessly with capillary blood is becoming increasingly important in connection with health screening in the event of complaints from athletes. Urea concentrations appear insufficient to claim valid inferences about the players’ acute stress levels, therefore urea isn’t an appropriate marker for the acute assessment of internal load but it could be an appropriate marker for assessing long-term  metabolic stress.Due to the players’ training schedule more standardized time points for sampling were not feasible. In addition to the blood sampling the session ratings of perceived exertion (sRPE) method seems a valid method of quantitating exercise training during a wide variety of types of exercise . In the Because of the high intra-individual variability of CK, urea, and CRP concentrations, testing of CK, urea and CRP should be performed as frequently as possible in order to (i) understand the kinetics of each variable during a season in relation to the given training load, and (ii) generate meaningful consequences for training and match tactics. Our data showed lower CK values toward the end of the half season. Therefore, we conclude that CK should be assessed with individual reference values compared to the midpoint of the U-19 first league half season. CK sensitively reflected soccer-specific loads during our observation suggesting this variable as a promising biomarker for internal load estimation when assessed frequently.The datasets generated for this study are available on request to the corresponding author.This study was approved by the ethics committee of the German Sport University . Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Staphylococcus aureus (S. aureus) infection in a rat model of infective endocarditis. The affinity of MH-ICG to S. aureus was determined and had a Km and Vmax of 5.4 μM and 3.0 X 10−6 μmol/minutes/108 CFU, respectively. MH-ICG had no detectable toxicity to mammalian cells at concentrations as high as 100 μM. The in vivo efficiency of MH-ICG in rats was evaluated using a right heart endocarditis model, and the accumulation of MH-ICG in the bacterial vegetations was 2.5 ± 0.2 times higher than that in the control left ventricular wall. The biological half-life of MH-ICG in healthy rats was 14.0 ± 1.3 minutes, and approximately 50% of injected MH-ICG was excreted into the feces after 24 hours. These data demonstrate that MH-ICG was internalized by bacteria with high specificity and that MH-ICG specifically accumulated in bacterial vegetations in a rat model of endocarditis. These results demonstrate the potential efficacy of this agent in the detection of infective endocarditis.Infectious endocarditis is a life-threatening disease, and diagnostics are urgently needed to accurately diagnose this disease especially in the case of prosthetic valve endocarditis. We show here that maltohexaose conjugated to indocyanine green (MH-ICG) can detect  Infectious endocarditis (IE) is a systemic infection in which bacteria form vegetations on the heart valves. The diagnosis of IE is made based on the modified Duke criteria in which the involvement of endocardium or supporting tissue is one of the major diagnostic criteria . Transthin vitro and in vivo and whether MH-ICG is accumulated in bacterial vegetations in a rat model of endocarditis.To develop novel diagnostic methods that detect the existence of bacteria in the tissue and organs, we have focused on the differences in sugar uptake by bacteria and mammalian cells –5. BacteAll animal protocols were approved by the Emory University Institutional Animal Care and Use Committee along with the guidelines and rules of Association for Assessment and Accreditation of Laboratory Animal Care International . All animal experiments were conducted in accordance with an approved protocol (PROTO201700806).2O (1:9) and filtered before purification using HPLC on a XBridge Prep C8 5 um OBD, 19 x 150 mm semi-prep column . The resulting MH-ICG was analyzed using mass spectrometry -5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS, CellTiter 96 AQueous One Solution Cell Proliferation Assay, Promega). The cells were incubated at 37°C for 60 minutes, and the formazan generated was measured at 490 nm with a microplate reader . The viability of MH-ICG treated cells was calculated as [absorbance of MH-ICG loaded cells]/[absorbance of control cells] (n = 4 /each concentration).CHO-K1 cells (ATCC CCL-61) cultured in F-12K medium with 10% FBS were used to evaluate the cytotoxicity of MH-ICG. On day -1, the cells were seeded in 96 well plates at density of 1 X 10Staphylococcus aureus  because S. aureus is one of the most commonly observed pathogens in patients with IE / [the mean intensity of the left ventricle free wall] was calculated for each slice. The average of the intensity ratio in 3 slices was used as the intensity ratio of the sample. In the control samples, the mean intensity in the right ventricle free wall was used instead of that of the vegetation.On POD 3, the IE rats were injected with 0.25 ml of 1 mM MH-ICG via the tail vein. Rats without catheterization and inoculation were used as controls. The rats were sacrificed 4 hours after injection of MH-ICG, and the hearts were sliced in three segments and scanned with an Male SD rats weighing 250 to 275 g were used for this experiment. The animals were anesthetized with 1–2% isoflurane and injected with 0.25 ml of 1 mM MH-ICG via the tail vein. At 15 and 30 minutes as well as 1, 2, 4, and 6 hours after injection of MH-ICG, 100 μl of blood was sampled with a heparinized syringe from the lateral tail vein opposite to the one used for injection of MH-ICG (n = 4 /each timepoint). The blood samples were centrifuged at 5000 X g for 10 minutes, and the plasma concentrations of MH-ICG were evaluated by a near infrared fluorescent microplate reader . The biological half-life of MH-ICG was calculated based on a two-compartment model with the following formula.Where Cn indicates the plasma concentration of MH-ICG in the sample n, tn indicates the sampled time point, and T indicates the biological half-life. Based on this formula, the biologil half-life in each animal and each time point was calculated.Male SD rats weighing 180 g to 190 g were injected with 0.25 ml of 1 mM MH-ICG via the tail vein and were housed in metabolic cages to collect feces and urine. The rats were sacrificed 24 hours after injection of MH-ICG, and the organs and tissue were harvested. The samples were homogenized with RIPA buffer using a TissueLyser II (Qiagen). The concentrations of MH-ICG in samples were evaluated with a fluorescent microplate reader  (n = 4).Analysis was performed with Prism statistical software (GraphPad Software). Student’s t-test was used for comparison of two groups, and P<0.05 was considered to be significant. All data are shown as mean ± standard error.S. aureus. The relationship between the concentrations of MH-ICG and the internalizing velocity of S. aureus is shown in S. aureus internalized MH-ICG even at low concentrations, and the uptake velocity plateaued at a concentration of 20 μM. For 108 CFU of S. aureus, the Km and Vmax calculated using a Lineweaver-Burk plot analysis were 5.4 μM and 3.0 X 10−6 μmol/minutes/108CFU, respectively.We first evaluated the kinetics of MH-ICG internalization by Escherichia. coli  and LamB mutant E. coli (JW-3996-1), which lacks maltoporin on the outer membrane [−7 μmol/minutes for 108 CFU of E. coli, respectively. The amount of MH-ICG internalized by 1 X 108 CFU of LamB mutant E. coli was 25.1 ± 4.4 pmol using 30 μM MH-ICG compared to 19.7 ± 1.1 pmol internalized with 2.5 μM MH-ICG. The uptake of MH-ICG by LamB mutant E. coli did not follow Michaelis-Menten kinetics consistent with the loss of specific maltoporin-mediated uptake. These results demonstrate that MH-ICG uptake by bacteria is highly specific and occurs via the maltodextrin transport system.We also evaluated the kinetics of MH-ICG in membrane . As showThe cytotoxicity of MH-ICG was evaluated in CHO-K1 cells. The MH-ICG showed no effects on cell growth or viability at concentrations of up to 100 μM for up to 3 days emphasizing the safety of MH-ICG for mammalian cells .S. aureus (8 CFU (n = 3).The rats developed right heart endocarditis two days after inoculation of . aureus . Histolo. aureus . Total bTo evaluate the accumulation of MH-ICG in the vegetations, the IE and control rats were injected with MH-ICG and sacrificed 4 hours after injection of MH-ICG. The pharmacokinetics of MH-ICG were evaluated in rats without infection. The MH-ICG rapidly cleared from the circulation with a pattern consistent with a dual mode of elimination . The halThe biodistribution studies revealed that approximately 50% of the injected MH-ICG was excreted within 24 hours with the majority of the product in the feces . The remin vitro assay system to demonstrate that MH-ICG is not taken up by mammalian cells and that uptake by bacteria largely occurs specifically through the maltodextrin transport system. The in vivo data confirm that systemically administered MH-ICG is effectively taken up by bacteria in our endocarditis model and that the free MH-ICG is effectively cleared from the circulation relatively quickly. The latter is important for both minimizing potential toxicity as well as decreasing background signal. In summary, these results support the concept that MH-ICG has the appropriate in vivo characteristics for localization of bacterial endocarditis.We report a novel and fluorescence-based imaging probe that can detect bacterial endocarditis lesions in a highly specific fashion. We used an 8 CFU, which is similar to that in the aortic valve endocarditis model [The model of endocarditis that we employed of these studies is somewhat unique in that it is a model of right heart endocarditis. Several previous studies of experimental endocarditis have used an aortic valve endocarditis model –23. We eis model –22.MH-ICG is our second generation of maltohexaose-based fluorescent probe; it has ICG as the probe molecule instead of IR-786 used in our first generation of maltohexaose-based fluorescent probes , 4. Both18F-MH is cleared primarily by the kidneys [The biodistribution studies were interesting in that approximately half of the injected MH-ICG was excreted into feces in 24 hours. This could reflect degradation of MH-ICG because ICG itself is known to be excreted into feces. Alternatively, the addition of ICG to MH might alter the clearance mechanism, i.e.,  kidneys .The limitation of MH-ICG in diagnosis of endocarditis is the permeability of the near infrared wavelength. Similar to ICG, the penetration of excitation and emission wavelength of MH-ICG is limited to a few centimeters of tissue, and endocarditis lesions could not be imaged through the chest wall with MH-ICG. However, visualization of the extent of bacterial infiltration could be assessed during valve replacement and other surgeries for infectious lesions such as orthopedic device infection; similar fluorescence imaging is done in certain settings , 28–31. In this study, we demonstrated that MH-ICG can determine the presence and extent of bacterial infection in the setting of endocarditis in a highly specific manner. MH-ICG has low cytotoxicity and is a viable candidate for further study of potential clinical applications; it may provide a unique diagnostic tool for bacterial endocarditis.S1 Fig(TIF)Click here for additional data file.S1 Data(DOCX)Click here for additional data file.


A bias-corrected dataset containing daily meteorological data over the Qinghai-Tibet Plateau has been generated using a trend-preserving bias correction, the Inter-Sectoral Impact Model Intercomparison Project (ISI-MIP) approach, with a high-quality gridded meteorological dataset based on ground observations (CN05.1). The dataset contains daily bias-corrected values of maximum/minimum near-surface air temperature, precipitation and mean near-surface wind speed from 15 models from the Fifth Phase of the Coupled Model Intercomparison Project (CMIP5) and a downscaled high-resolution dataset (NEX-GDDP), based on CMIP5 models, over the Qinghai-Tibet Plateau (QTP) during 1986–2095. This dataset provides an important reference for the study of future climate change and its impacts in the Qinghai-Tibet Plateau region. Specifications table•This dataset includes daily maximum/minimum near-surface air temperature, precipitation and mean near-surface wind speed from 15 models under the RCP4.5 and RCP8.5 scenarios. Its accuracy is further improved by being bias-corrected using local observation data (CN05.1), containing more observation gauges.•This dataset is of great value to researchers who study the impacts and risks of climate change in the QTP. Its spatial and temporal resolution allows potential users to develop their impact or risk assessment models at a higher resolution in this region.•For further insights and development of experiments, for example, it can be used to predict future extreme weather events, crop yields, etc. Furthermore, using data from 15 models enables multi-model ensemble analysis, which is considered to be essential in climate projection and climate risk analysis.1In the following, “NEX-GDDP/GCMs before bias correction” refers to the original data that we downloaded from the relevant websites and “NEX-GDDP/GCMs after bias correction” refers to the data that we have bias-corrected by applying the ISI-MIP approach.The raw dataset contains daily maximum/minimum near-surface air temperature, precipitation and mean near-surface wind speed from 15 models see , which wFor analyzed data, we processed the daily bias-corrected data into annual, winter  and summer  values and then calculated the differences between NEX-GDDP/GCM data before/after bias correction and CN05.1  to show the improvement from applying the bias correction process. For mean values, we use annual, winter and summer average of Tmax, Tmin, Pr, and Wind. For climate extreme values, we use the 95th percentile of Tmax, the 5th percentile of Tmin, the 95th percentile of Pr and the 95th percentile of Wind. The trend values are represented by the slope of the unary linear regression of the observed/multi-model time series.The rest of this section shows the differences between the NEX-GDDP/GCM data before/after bias-correction and the reference data.1.1As can be seen from 1.2For the future period, the bias correction process adjusts the values to better relate to historical values a and 4a,2The models to be bias-corrected contain two sets of simulated data. Fifteen models were selected, with data from a training period (1986–2005), a validation period (1966–1985) and a future period (2006–2095/2099/2100) under two Representative Concentration Pathway (RCP) scenarios, 4.5 and 8.5. Tmax, Tmin and Pr were obtained from NEX-GDDP and Wind was obtained from CMIP5. Tmax and Tmin are at a height of 2 meters and Wind is at a height of 10 meters. Near-surface wind is important to climate change in terms of the carbon exchange, water transport, evapotranspiration, etc., but NEX-GDDP does not contain this variable. We aim to provide a dataset containing interpolated and bias-corrected wind speeds that are convenient to use in research. NEX-GDDP is a statistical downscaling dataset based on CMIP5. In our dataset, Tmax, Tmin and Pr (from NEX-GDDP) and Wind (from CMIP5) come from the same 15 models and the same ensemble (r1i1p1).The reference dataset is CN05.1 The region studied extends from 73.25°E to 104.75°E and from 26°N to 39.75°N. CMIP5 data were interpolated into a 0.25° × 0.25° grid using a nearest-neighbor interpolation method. We divided the historical period (1966–2005) into two parts: data for 1986–2005 were used for training and data for 1966–1985 were used for validation. A trend-preserving bias correction, the Inter-Sectoral Impact Model Intercomparison Project (ISI-MIP) approach 1) Correction for monthly meanC was used to bias-correct temperature, which was the difference of 20-year monthly mean temperature of CN05.1 and GCMs:i represented year, Following the algorithm, a constant offset c was used to bias-correct precipitation and wind, which was the ratio of 20-year monthly mean precipitation/wind of CN05.1 and GCMs. Take precipitation as an example,Multiplicative correction factor 2) Correction for daily variabilityIn the second step, daily variability was corrected by mapping daily residuals. For temperature,i, day j was simply the difference between the daily value and its corresponding monthly value.The residual temperature For precipitation and wind, the daily residual was the ratio between daily value and corresponding monthly value. Take precipitation as an example,TCN05.1) and GCMs data (ΔGCMT).The variability of daily residuals of the GCMs was adjusted by using some transfer functions. For temperature, we had the functionτ were the parameters of the function. Superscript The daily variability of precipitation and wind was adjusted using nonlinear regression, see Ref .f(ΔGCMT) and Finally, the bias-corrected data of temperature was:After that, the correction parameters for the training period were then derived. They were validated for the period 1966–1985. Validation results indicated that parameters based on the training dataset worked fairly well for the validation dataset, and therefore suggested plausible applicability to the future period. These parameters were then applied to bias-correct the future period (2006–2095/2099/2100). More details of the bias correction approach can be found in Ref The authors declare that they have no known competing financial interests or personal relationships which have, or could be perceived to have, influenced the work reported in this article.